Changes in Blood Parameters After Intramuscular Testosterone Ester Injections - Implications for Anti-Doping

Changes in Blood Parameters After Intramuscular Testosterone Ester Injections : Implications for Anti-Doping / Sara Amalie Solheim, Jakob Mørkeberg, Yvette Dehnes, Ingunn Hullstein, Anders Juul, Emmie N. Upners, Nikolai Baastrup Nordsborg. - (Drug Testing and Analysis (2020) 20 April)

  • PMID: 32307878.
  • DOI: 10.1002/dta.2803


Testosterone treatment stimulates red blood cell production and alters iron homeostasis. Thus, we investigated whether the 'hematological module' of the Athlete Biological Passport (ABP) used by the World Anti-Doping Agency (WADA) can be used to indicate misuse of testosterone. Nineteen eugonadal men received intramuscular injections of either 250 mg Sustanon®, a blend of four testosterone esters, or placebo on Day 0 and 21 in a randomized, placebo-controlled double-blind design. Urine samples and blood samples for the assessment of steroidal and hematological biomarkers of the ABP were collected twice pre-treatment, at least 5 days apart, and on days 1, 3, 5, 10 and 14 post-injections. The steroidal profile was flagged suspicious in all Sustanon®-treated subjects, while the hematological profile was flagged suspicious in 6 out of 9 subjects. When considering both sensitivity and specificity, RET% appeared as the best marker of the hematological module for implying testosterone ester misuse. Atypical blood passport samples were used to select time points for further isotope-ratio mass spectrometry (IRMS) analysis of testosterone and its metabolites in simultaneously collected urine. The RET% and OFF-Score could, in addition to the T/E ratio, help identify suspicious samples for more targeted IRMS testing. The present results demonstrate that unexpected fluctuations in RET% can be indicative of testosterone doping if samples are collected 3-10 days after injection. From an anti-doping perspective, the hematological and steroidal modules of the ABP should complement each other when planning targeted follow-up testing and substantiating likely misuse of testosterone.


Research / Study
20 April 2020
Dehnes, Yvette
Hullstein, Ingunn R.
Juul, Anders
Mørkeberg, Jakob
Nordsborg, Nikolai Baastrup
Solheim, Sara Amalie
Upners, Emmie N.
Adverse Analytical Finding / presence
Other organisations
Anti-Doping Denmark (ADD)
Københavns Universitet - University of Copenhagen (UCPH)
Oslo universitetssykehus HF - Oslo University Hospital
Oslo, Norway: Norwegian Doping Control Laboratory
Analytical aspects
Mass spectrometry analysis
Testing method development
Doping classes
S1. Anabolic Agents
T/E ratio (testosterone / epitestosterone)
Medical terms
Hematocrit levels
Athlete Biological Passport (ABP)
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Date generated
22 April 2020
Date of last modification
7 September 2020
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